Sugar-conjugated lipid nanoparticles and methods of use therefor

Abstract

Provided are compositions that include compositions of galactosyl-conjugated lipid nanoparticles (LNPs) encapsulating one or more active agents. In some embodiments, the galactosyl-conjugated LNPs have a lipid component having D-Lin-MC3-DMA, ALC-0315 and SM-102, cholesterol. DSPC and DOPE, and DMG-2000-PEG. In some embodiments, the GalNAc-conjugated LNP has one or more galactosyl moieties bioconjugated to cholesterol present with a lipid component of the GalNAc-conjugated LNP. Also provided are methods for treating and/or preventing diseases, disorders, and/or conditions associated with undesirable PCSK9 gene expression, optionally a cardiovascular disease, disorder, or condition, including but not limited to atherosclerosis and/or thrombosis, and sepsis, septic shock, cytokine storm, or sequelae thereof.

1 . A method for treating and/or preventing a disease, disorder, or condition associated with undesirable proprotein convertase subtilisin/kexin type 9 (PCSK9) gene expression, the method comprising administering to a subject in need thereof an effective amount of a comprising a sugar-conjugated lipid nanoparticle (LNP) and an active agent encapsulated therein, optionally wherein the sugar is a hexose monosaccharide, wherein the active agent modulates the PCSK9 gene expression to thereby treat and/or prevent the disease, disorder, or condition. 
     
     
         2 . The method of  claim 1 , wherein the sugar is selected from the group consisting of mono-, di-, and triannary galactosyl, N-acetyl galactosamine (GalNac), glucose, N-acetyl glucosamine (GluNac), mannose, trehalose, fucoidan, pyranose, and furanose. 
     
     
         3 . The method of  claim 1 , wherein the LNP is a galactosyl-conjugated LNP, and the galactosyl-conjugated LNP further comprises a lipid component comprising about 5-10% cholesterol. 
     
     
         4 . The method of  claim 1 , wherein the LNP, optionally the galactosyl-conjugated LNP, comprises a lipid component comprising one or more of hydrolyzable ionizable lipids, optionally selected from the group consisting of D-Lin-MC3-DMA, ALC-0315, and SM-102; cholesterol and/or an analog thereof, optionally β-sitosterol; and optionally one or more helper lipids, optionally selected from the group consisting of DSPC, DOPE, and DMG-2000-PEG, further optionally two, three, or all four of D-Lin-MC3-DMA, cholesterol, DSPC, and DMG-2000-PEG. 
     
     
         5 . The method of  claim 1 , wherein the LNP is a galactosyl-conjugated LNP that comprises one or more galactosyl moieties bioconjugated to cholesterol present with a lipid component of the galactosyl-conjugated LNP. 
     
     
         6 . The method of  claim 1 , wherein the active agent is a nucleic acid, optionally an inhibitory nucleic acid, further optionally an siRNA. 
     
     
         7 . The method of  claim 6 , wherein the inhibitory nucleic acid inhibits a biological activity of a PCSK9 gene product, optionally a human PCSK9 gene product, and further optionally wherein the inhibitory nucleic acid is an inclisiran derivative lacking a GalNAc moiety. 
     
     
         8 . The method of  claim 1 , wherein the LNP, optionally the galactosyl-conjugated LNP, targets the active agent to a cell, tissue, or organ of interest, optionally wherein the cell tissue, or organ of interest is selected from the group consisting of vasculature or a vasculature cell, optionally a vascular endothelial cell, a muscle cell, a fibroblast, and/or a non-vascular endothelial cell. 
     
     
         9 . The method of any one of  claims 1 - 8 , wherein the LNP comprises a ratiometric combination of ionizable lipids. 
     
     
         10 . The method of  claim 9 , wherein the LNP comprises a ratiometric combination of ionizable lipids with a specific dissociation constant value of between about 6.05 and about 6.44. 
     
     
         11 . The method of  claim 1 , wherein the LNP comprises one or more mono-, di-, and/or ternary hydrophobic fatty acids and/or one or more hydrophilic polyethylene glycols conjugated to the sugar moiety onto cholesterol at its head or tail and/or onto a polyethylene glycol (PEG) moiety. 
     
     
         12 . The method of  claim 11 , wherein at least one of the one or more fatty acids and/or the PEG moiety comprises a chain length of 1 to 15 carbons. 
     
     
         13 . The method of  claim 1 , wherein the disease, disorder, or condition is selected from the group consisting of an inflammatory disease, disorder, or condition; a cardiovascular disease, disorder, or condition, optionally atherosclerosis and/or thrombosis; and sepsis, septic shock, cytokine storm, or sequelae thereof. 
     
     
         14 . The method of  claim 13 , further comprising administering to the subject in need thereof an effective amount of a second active agent that is appropriate for treating the disease, disorder, or condition. 
     
     
         15 . The method of  claim 14 , wherein the disease, disorder, or condition is:
 (i) an inflammatory disease, disorder, or condition, and the second active agent is an anti-inflammatory agent, optionally a nonsteroidal anti-inflammatory drug (NSAID), a steroid, acetaminophen or other COX-3 inhibitor, a 5-lipoxygenase inhibitor, a leukotriene receptor antagonist, a leukotriene A4 hydrolase inhibitor, an antihistaminic, a histamine 2 receptor antagonist, a phosphodiesterase-4 antagonist, a cytokine antagonist, a CD44 antagonist, an antineoplastic agent, a 3-hydroxy-3-methylglutaryl coenzyme A inhibitor (e.g., a statin), an estrogen, an androgen, an antiplatelet agent, an antidepressant, a Helicobacter pylori inhibitor, a proton pump inhibitor, a thiazolidinedione, or a dual-action compound;   (ii) a cardiovascular disease, disorder, or condition, optionally atherosclerosis, and the second active agent is an acyl-coenzyme A: cholesterol acyltransferase (ACAT) inhibitor, including but not limited to a selective inhibitor of ACAT-1, ACAT-2 as well as dual inhibitors of ACAT-1 and ACAT-2; an alpha-adrenergic blocking drug (alpha-blocker), an alpha/beta blocker, an angiotensin-converting enzyme (ACE) inhibitor, an aldosterone antagonist, an angiotensin II receptor antagonist, an anti-arrhythmic, an anticoagulant, an antiplatelet agent, an apolipoprotein A-1 (apoA-1) mimetic, a beta-blocker, a bile acid sequestrant, a calcium-channel blocker, an ApoB cholesteryl ester transfer protein (CETP) inhibitor, a cholesterol absorption inhibitor, a diuretic, a dyslipidemia agent, an endothelin receptor antagonist, a fibrate, a 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor, an LCAT activator, an LDL receptor inducer, a lipase inhibitor, a lipoprotein-associated phospholipase A2 (Lp-PLA2) inhibitor, a microsomal triglyceride transfer protein (MTP) inhibitor, a platelet aggregation inhibitor, a PPAR agonist and/or activator, optionally a PPARγ agonist, PPARα agonist, and/or a PPAR dual α/γ agonists, a second PCSK9 antisense nucleic acid or RNAi, a squalene epoxidase inhibitor, a squalene synthetase inhibitor, a thrombolytic, a thyroid receptor beta activator, or any combination thereof;   (iii) a cardiovascular disease, disorder, or condition, and the second active agent targets a vasculature cell, a muscle cell, a fibroblast, and/or an endothelial cell, optionally a vascular endothelial cell, wherein the second active agent binds to a target selected from the group consisting of LOX-1, a selectin, optionally a P-selectin or an E-selectin, an integrins, and a neuropilin; and/or   (iv) sepsis, septic shock, or cytokine storm and the second active agent is an antibiotic, a complement inhibitor, optionally a complement inhibitor selected from the group consisting of a C5a inhibitor, a C5aR inhibitor, a C3 inhibitor, optionally wherein the C3 inhibitor is Compstatin, a Compstatin analog, a Compstatin peptidomimetic, or a Compstatin derivative, a factor D inhibitor, a factor B inhibitor, a C4 inhibitor, a C1q inhibitor, or any combination thereof.   
     
     
         16 . A method for targeting an active agent to a hepatocyte, the method comprising contacting the hepatocyte with a composition comprising an active agent encapsulated by a sugar-conjugated lipid nanoparticle (LNP), optionally a hexose monosaccharide-conjugated LNP, further optionally an N-acetylgalactosamine (GalNAc) conjugated lipid nanoparticle (LNP), wherein the sugar-conjugated LNP comprises a lipid component comprising ionizable aminolipid ALC-0315 (6-((2-hexyldecanoyl)oxy)-N-(6-((2-hexyldecanoyl)oxy)hexyl)-N-(4-hydroxybutyl)hexan-1-aminium) and one or more of D-Lin-MC3-DMA, cholesterol, DSPC, DOPE, and DMG-2000-PEG, optionally two, three, four, or all five of D-Lin-MC3-DMA, cholesterol, DSPC, and DMG-2000-PEG, further optionally wherein the lipid component comprises cholesterol with one or more GalNAc moieties conjugated thereto, optionally with 25% of the cholesterol being galactosyl-cholesterol, whereby the active agent is targeted to the hepatocyte. 
     
     
         17 - 22 . (canceled)